The main roles of the Protein Production and Chemistry Core (PCC) are to produce modified proteins for the Bridge and Pilot Projects, and provide advice, reagents, methods development and assistance to investigators inside and outside the Membrane Protein Structural Dynamics Consortium, MPSDC. The PCC brings together extensive experience in producing labeled integral membrane proteins in prokaryotic and eukaryotic cells, cell- free, or synthetically, as well as chemical synthesis of novel spectroscopic probes. We have prepared homogeneous functional proteins for use in studies of structure, dynamics and mechanism. The Core members already have a wide range of systems in place for producing proteins in prokaryotic and eukaryotic systems as well as cell-free and synthetic approaches coupled with expertise in membrane protein folding. In addition, the PCC members have collaborated to develop new expression approaches with a focus on incorporation of spectroscopic probes for monitoring protein dynamics in vitro and in vivo. To serve the MPSDC and to provide service to any interested investigator, the PCC will carry out these specific aims: (1) collaborate with MPSDC members to develop and carry out membrane protein expression, solubilization, purification, reconstitution and characterization of homogeneous, folded and functional proteins; (2) service the community by providing advice, protocols, tools and reagents on aspects of membrane protein production, including expression, solubilization, purification, folding, modification and characterization in order to generate homogeneous functional proteins using bacterial, fungal, insect and mammalian cell, cell-free and synthetic systems. An important part of the service is providing a web-based resource for investigators to find reagents and tools for membrane protein production, and useful information with links to sources of reagents; (3) develop new efficient and optimized reagents for site-directed labeling with spectroscopic probes, with emphasis on specific properties to enhance their utility in structural dynamics. The probes will be designed for chemical modification as well as direct incorporation during protein synthesis as unnatural amino acids.